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组织培养和农杆菌介导的油料作物向日葵遗传转化。

Tissue culture and Agrobacterium-mediated genetic transformation of the oil crop sunflower.

机构信息

Xinjiang Key Laboratory of Biological Resources and Genetic Engineering, College of Life Science and Technology, Xinjiang University, Urumqi, XinJiang, China.

Key Laboratory of Microbial Resources Protection, Development and Utilization, College of Biological Sciences and Technology, Yili Normal University, Yining, XinJiang, China.

出版信息

PLoS One. 2024 May 9;19(5):e0298299. doi: 10.1371/journal.pone.0298299. eCollection 2024.

Abstract

Sunflower is one of the four major oil crops in the world. 'Zaoaidatou' (ZADT), the main variety of oil sunflower in the northwest of China, has a short growth cycle, high yield, and high resistance to abiotic stress. However, the ability to tolerate adervesity is limited. Therefore, in this study, we used the retention line of backbone parent ZADT as material to establish its tissue culture and genetic transformation system for new variety cultivating to enhance resistance and yields by molecular breeding. The combination of 0.05 mg/L IAA and 2 mg/L KT in MS was more suitable for direct induction of adventitious buds with cotyledon nodes and the addition of 0.9 mg/L IBA to MS was for adventitious rooting. On this basis, an efficient Agrobacterium tumefaciens-mediated genetic transformation system for ZADT was developed by the screening of kanamycin and optimization of transformation conditions. The rate of positive seedlings reached 8.0%, as determined by polymerase chain reaction (PCR), under the condition of 45 mg/L kanamycin, bacterial density of OD600 0.8, infection time of 30 min, and co-cultivation of three days. These efficient regeneration and genetic transformation platforms are very useful for accelerating the molecular breeding process on sunflower.

摘要

向日葵是世界四大油料作物之一。中国西北地区的主要油用向日葵品种“早大头”(ZADT)具有生长周期短、产量高、抗非生物胁迫能力强等特点,但耐逆境能力有限。因此,本研究以保留系骨干亲本 ZADT 为材料,建立其组织培养和遗传转化体系,通过分子育种提高其抗性和产量。在 MS 培养基中,0.05mg/L IAA 和 2mg/L KT 更适合子叶节不定芽的直接诱导,而在 MS 培养基中添加 0.9mg/L IBA 则更适合不定根的诱导。在此基础上,通过卡那霉素筛选和转化条件优化,建立了高效的农杆菌介导的 ZADT 遗传转化体系。在 45mg/L 卡那霉素、OD600 为 0.8、侵染时间 30min、共培养 3d 的条件下,PCR 检测阳性苗率达到 8.0%。这些高效的再生和遗传转化平台对于加速向日葵的分子育种进程非常有用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/faf4/11081250/c91fe0d9b3fe/pone.0298299.g001.jpg

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